Detection and characterization of ZnT8 autoantibodies could help to screen latent autoimmune diabetes in adult-onset patients with type 2 phenotype

Abstract

Autoantibodies to zinc transporter 8 (ZnT8A) constitute an additional marker of autoimmune diabetes, complementing those already used in diagnosis support. ZnT8A could also be found in latent autoimmune diabetes of adults (LADA). The aim of this study was to evaluate the prevalence of ZnT8A in adult-onset diabetic patients in Argentinian population. A total of 271 patients diagnosed for diabetes at mean age 53.4 ± 10.9, body mass index ≤ 30, without insulin treatment for the first year of disease, and initially classified as type 2 diabetic patients were tested for ZnT8A using cDNA plasmids encoding the C-terminal domains (aa 268-369) carrying 325Arg, 325Trp, and a dimeric cDNA construct carrying both 325Arg and 325Trp (ZnT8 Arg-Trp325). We also analyzed proinsulin autoantibodies (PAA), glutamic acid decarboxylase autoantibodies (GADA), and protein tyrosine phosphatase IA-2 autoantibodies (IA-2A). A subset of 101 patients was followed during 6 years in order to analyze insulin requirement. Out of the 271 patients, 22.1% presented at least one humoral marker, 2.6% were PAA+, 12.5% were GADA+, 3.3% were IA-2A+, and 10.7% were ZnT8A+. Among the latter, 7.0% were ZnT8A-Arg325, 51.7% were ZnT8A-Trp325, and 62.1% were ZnT8A-Arg-Trp325. Furthermore, the prevalence of autoantibodies in the group of patients treated with insulin (n = 18) was 55.6%. These results demonstrated that a significant proportion of autoimmune adult-onset diabetic patients presented ZnT8A as the only humoral marker. Between them, the higher prevalence was for ZnT8A-Trp325. We suggest that screening for LADA patients, best performed with a minimal set of marker determination, must include at least the screening of GADA and ZnT8A-Arg-Trp325.

Zinc transporter 8 autoantibodies in patients with type 1 diabetes from a multiethnic population and their first degree relatives

“…Moreover, Trabucchi and cols. have shown that a significant proportion of autoimmune adult-onset diabetic patients present ZnT8A as the only humoral marker (32).…”

Characterization of zinc transporter 8 (ZnT8) antibodies in autoimmune diabetic patients from Argentinian population using monomeric, homodimeric, and heterodimeric ZnT8 antigen variants

“…Adults (LADA). In a previous report (11,19) we have proved that patients with adult-onset DM initially identified as 'marker negative', based on GADA and IA-2A screening, were reclassified as autoantibody positive when tested for ZnT8A. In the present study, we have included homodimer ZnT8 probes in order to improve the differential characterization of ZnT8A in type 1 and non-obese adult-onset diabetic patients.…”

“…ZnT8A were assessed by radioligand binding assay (RBA) as previously described (11). Briefly, cDNA coding for ZnT8-Arg325, ZnT8-Trp325, ZnT8-Arg-Arg325, ZnT8-TrpTrp325, and ZnT8-Arg-Trp325 were transcribed and translated using a rabbit reticulocyte lysate system (Promega Corporation) and …”

“…ZnT8A have been detected in more than 60% of patients with type 1 DM (9, 10) and in more than 10% of adult-onset diabetic patients (11).…”

“…In a previous work, we have characterized ZnT8A prevalence in Argentinian non-obese adult-onset diabetic patients only using monomer and hetero-dimer variants of the ZnT8 C-terminal domain (11). In the present study, the role of amino acid at position 325 and dimeric construction were analyzed in order to gain further knowledge of the structure of ZnT8 epitopes and ZnT8A recognition.…”